2-aminopyridine-based selective neuronal nitric oxide synthase inhibitors

ABSTRACT

Aminopyridine compounds, as can be used in conjunction with methods for modulation of nitric oxide synthase activity.

This application is a divisional of and claims priority to and thebenefit of application Ser. No. 14/840,801 filed Aug. 31, 2015 andissued as U.S. Pat. No. 9,416,106 on Aug. 16, 2016, which was adivisional of and claimed priority to and the benefit of applicationSer. No. 14/199,599 filed Mar. 6, 2014 and issued as U.S. Pat. No.9,120,750 on Sep. 1, 2015, which claimed priority to and the benefit ofprior provisional application Ser. No. 61/774,147 filed Mar. 7,2013—each of which is incorporated herein by reference in its entirety.

This invention was made with government support under GM049725 awardedby the National Institutes of Health. The government has certain rightsin the invention.

BACKGROUND OF THE INVENTION

Neuronal nitric oxide synthase (nNOS) catalyzes the oxidation ofL-arginine to L-citrulline in the central nervous system, generatingnitric oxide (NO), a critical neurotransmitter. Significant research hasimplicated the overexpression of nNOS—and overproduction of NO—invarious neurological diseases, including Parkinson's, Alzheimer's, andHuntington's diseases, as well as neuronal damage due to stroke,cerebral palsy and migraine headaches. Inhibiting endothelial nitricoxide synthase (eNOS) and inducible nitric oxide synthase (iNOS) is,however, undesirable, because these isozymes are responsible formaintaining crucial body function. Thus, selective inhibition of nNOSover its closely related isoforms, eNOS and iNOS, can provide apromising strategy in developing therapeutics for the treatment ofneurodegenerative diseases.

Many organic scaffolds have been investigated over the past decade. Somecompounds exhibit good potency and high selectivity, but suffer frompoor bioavailability, thereby hampering their therapeutic potential. Yetother compounds, while promising, are limited by synthetic challengeand/or low yield. As a result, there remains an on-going concern in theart to provide an efficient approach toward a facile synthesis of a widerange of such compounds, with structures optimized for enhancedpharmacological effect.

SUMMARY OF THE INVENTION

In light of the foregoing, it is an object of the present invention toprovide compounds, compositions and related methods of use for theselective inhibition of neuronal nitric oxide synthase, therebyovercoming various deficiencies and shortcomings of the prior artincluding those outlined above. It would be understood by those skilledin the art that one or more aspects of this invention can meet certainobjectives, while one or more other aspects can meet certain otherobjectives. Each objective may not apply equally, in all its respects,to every aspect of this invention. As such, the following objects can beviewed in the alternative with respect to any one aspect of thisinvention.

It is an object of the present invention to provide one or more smallmolecule and/or non-peptide compounds exhibiting selective nNOSinhibition over other enzyme isoforms and providing improved membranepermeability and bioavailability.

It can be another object of the present invention to provide one or moresuch compounds incorporating moieties found in natural amino acids andother natural products, to optimize pharmacological profile.

It can be another object of the present invention, alone or inconjunction with one or more of the preceding objectives, to provide aversatile synthetic route for incorporation of such naturally-derivedmoieties into compounds comprising one or more aminopyridine moietiesshown effective in promoting selective nNOS inhibition.

It can be another object of the present invention to provide one or moresuch compounds for in vitro use and study under conditions promotingnitric oxide production, indicative of one or more mammalian diseasestates.

Alternatively, it can also be an object of the present invention toprovide one or more such compounds enabling in vivo treatment of suchdisease states.

Other objects, features, benefits and advantages of the presentinvention will be apparent from this summary and the followingdescriptions of certain embodiments of such compounds, compositionsand/or methods and will be readily apparent to those skilled in the arthaving knowledge of the synthetic techniques described herein. Suchobjectives, features, benefits and advantages will be apparent from theabove as taken into conjunction with the accompanying examples, data,figures and references incorporated herein, together with all reasonableinferences to be drawn therefrom.

In part, the present invention can be directed to compounds of a formula

wherein R¹ can be selected from 6-amino-4-methylpyridin-2-yl and6-aminopyridin-2-yl moieties; X can be selected from CH₂O and CH₂CH₂moieties; Y can be selected from O and CH₂ moieties; and n can beselected from 1 and 2, and salts, hydrates and solvates thereof.

In part, the present invention can also be directed to compounds of aformula

wherein X can be selected from N and CH; L can be selected fromO(CH₂)_(n) and CH₂NH(CH₂)_(n) moieties, and where n can be selected from1-3; and R¹ can be selected from amino, pyridinyl, piperidinyl, phenyl,fluorophenyl and chlorophenyl moieties, and salts, hydrates and solvatesthereof.

In part, the present invention can also be directed to compounds of aformula

wherein R¹ can be selected from 6-aminopyridin-2-yl,6-amino-4-methylpyridin-2-yl, phenyl, fluorophenyl and chlorophenylmoieties; R² can be selected from H, methyl and ethyl moieties; and Lcan be selected from CH₂O, CH₂CH₂, CH₂N, CH₂O(CH₂)₂O, (CH₂)₅ andCH₂N(CH₂)₂N moieties, and salts, hydrates and solvates thereof.

In part, the present invention can also be directed to compounds of aformula

together with salts, hydrates and solvates thereof.

Regardless, the compounds of this invention are without stereochemicallimitation. As illustrated and discussed below, such compounds and/ortheir intermediates are available as racemic mixtures from which isomerscan be resolved, or are diastereomers from which the correspondingenantiomers can be separated. Accordingly, any stereocenter can be (S)or (R) with respect to any other stereocenter(s). As a separateconsideration, various compounds can be present as an acid salt, eitherpartially or fully protonated. In certain such embodiments, thecounterion(s) can be a conjugate base of a protic acid. Further, it willbe understood by those skilled in the art that any one or more thecompounds of this invention can be provided as part of a pharmaceuticalcomposition comprising a pharmaceutically-acceptable carrier componentfor use in conjunction with a treatment method or medicament.

In part, the present invention can also be directed to a methodinhibiting, modulating or otherwise affecting a nitric oxide synthase.Such a method can comprise providing a compound of this invention,whether or not part of a pharmaceutical composition, and administeringan effective amount of such a compound for contact with a nitric oxidesynthase, such compounds including but not limited to those illustratedby the following examples, referenced figures and/or accompanyingsynthetic schemes. In certain such embodiments, such a compound and/orcombination thereof can be present in an amount at least partiallysufficient to selectively inhibit neuronal nitric oxide synthase overinducible and endothelial isoforms.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1. Chemical structures of representative, non-limiting inhibitorcompounds, in accordance with certain embodiments of this invention.

FIG. 2. Schematic chemical structures of representative, non-limitingselective nNOS inhibitors—also in accordance with this invention.

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS

The following illustrates preparation and use of various NOS inhibitorcompounds, in accordance with certain non-limiting embodiments of thisinvention. Using well-known synthetic techniques, as would be understoodby those skilled in the art made aware of this invention, representativecompounds 1-21 were prepared, as shown. Structural analogues of suchcompounds can be prepared using such techniques or straight-forwardvariations thereof, such analogous compounds limited only by commercialor synthetic availability of corresponding starting materials andreagents, such techniques, variations, starting materials and reagentsas would also be understood by those skilled in the art and made awareof this invention. Many of the present compounds involve aminopyridinecomponent 22, and its synthesis from available starting materials isdescribed in Scheme 1. The synthesis of compound 1 starts from thecommercially available resorcinol and reaction with N-protected bromideintermediate, generating compound 23. Coupling via an ether synthesisprovided compound 24. The final product, compound 1, was obtained in ayield of 74% after deprotection.

With reference to scheme 2, ether synthesis was used to couple apyridinyl fragment with resorcinol, and a second ether synthesisconnected an aminopyridine component to the backbone. Compound 2 wasobtained in a yield of 89% after removal of the dimethylpyrroleprotecting group.

The structure and synthesis of compound 3 are similar to compound 2. Asshown in scheme 3, Mitsunobu reaction linked a piperidinyl compound tothe resorcinol linker, which was followed by an ether synthesis togenerate compound 28. After the respective removal of the two protectinggroups, the final compound 3 was obtained in a good yield.

With reference to scheme 4, ether synthesis was used to first couple anaminopyridine component to the aldehyde linker. Reductive amination thenproduced compound 31. Compound 4 was obtained successfully afterremoving the protecting group.

With reference to scheme 5, ether synthesis was first used to connectthe aminopyridine component to the pyridinyl linker, then the methylester was reduced with Dibal-H to the aldehyde. Reductive aminationproduced compound 34. (Compounds incorporating a chlorophenyl moiety areavailable through use of a corresponding chloro-substituted amine.)Compound 5 was obtained successfully after removing the protectinggroup.

Compounds 6-9 were prepared using the same synthetic route, from theirrespective starting materials. For instance, as shown in Scheme 6, thecommercially available proline analogue was reduced with LiBH₄ togenerate diol 35. Then, a one-step ether synthesis linked twoaminopyridine components to produce compound 36. The final compound 6was obtained after the respective removal of the dimethylpyrrole and Bocprotecting groups. Compounds 7-9 were synthesized via the same methodwith similar yields.

Compounds 10-11 were prepared using the same synthetic route, from theirrespective starting materials. As shown in scheme 7, a firstintermediate compound 38 was prepared from 22 after ether synthesis withsubsequent bromination. Second fragment compound 40 was synthesizedthrough an ether synthesis and subsequent reduction with LiBH₄.Compounds 38 and 40 were coupled to produce 41, with compound 10obtained after deprotection. Compound 11 was synthesized via the samemethod with similar yields.

Compounds 12-13 were prepared using the same synthetic route, and shareintermediate 40 with compounds 10-11. As shown in scheme 8, secondintermediate compound 44 was prepared from 3-fluorophenylmethanol, withsubsequent breakage of C═C bond, reduction to alcohol 43 andbromination. Compounds 44 and 40 were coupled via ether synthesis toproduce 45, with compound 12 then obtained after deprotection. Compound13 was synthesized via the same method in similar yield.

With reference to scheme 9, the diol starting material was firstdimethylpyrrole protected, then coupled with two aminopyridinecomponents via a one-step reaction to generate 47. The final compound 14was obtained after the removal of the three protecting groups under thesame conditions. Compound 15 was synthesized via the same method insimilar yield.

With reference to Scheme 10, the synthesis of compound 16 started withan aminobutyric acid analogue. After reduction of the acid to alcoholand amino group protection, compound 49 was generated in good yield. Twoaminopyridine components were coupled thereto via a one-step ethersynthesis in a yield of 68%. Final compound 16 was obtained after theremoval of the three protecting groups under the same conditions.Compound 17 was synthesized via the same method in similar yield.

As shown in scheme 11, the synthesis of compound 18 followed the samesynthetic method as used for compounds 16-17, from the correspondingaminobutyric acid starting material.

The compounds, compositions and/or methods of the present invention cansuitably comprise, consist of, or consist essentially of any of theaforementioned moieties or substituents and/or functional groupsthereof. Each such compound or moiety/substituent/functional groupthereof is compositionally distinguishable, characteristicallycontrasted and can be practiced in conjunction with the presentinvention separate and apart from another. Accordingly, it should alsobe understood that the inventive compounds, compositions and/or methods,as illustratively disclosed herein, can be claimed, practiced orutilized in the absence of any one compound,moiety/substituent/functional group which may or may not be disclosed,referenced or inferred herein, the absence of which may or may not bespecifically disclosed, referenced or inferred herein.

With reference to scheme 12, compounds 19-21 were prepared using thesame synthetic route, from their respective starting materials. Afterprotection of the amino group, the methyl ester was reduced with LiBH₄to the alcohol. The two aminopyridine components were coupled via aone-step ether synthesis in a yield of 73%. The final compound 19 wasobtained after the removal of the three protecting groups under the sameconditions. Compounds 20-21 were synthesized via the same method withsimilar yields.

Demonstrating utility of this invention, NOS inhibition assays ofrepresentative compounds 1-21 were undertaken, and the results aresummarized in Table 1, below. All NOS isoforms were expressed andpurified as described in U.S. Pat. No. 7,470,790 and the literaturereferences cited therein, each of which is incorporated herein byreference in its entirety. Nitric oxide formation from NOS was monitoredusing literature techniques. Again, reference is made to theaforementioned '790 patent and the references cited therein. (See, e.g.,Hevel, J. M.; Marietta, M. A. Nitric Oxide Synthase Assays. MethodsEnzymol. 1994, 133, 250-258).

TABLE 1 Inhibition of NOS isozymes by synthetic inhibitors.^(a) Ki [nM]Selectivity^(b) Compounds nNOS eNOS iNOS e/n i/n 1 60 18018 8472 298 1402 616 62832 27104 102 44 3 117 27963 17124 239 146 4 40 14414 9914 260147 5 140 7114 2862 74 21 6 26 2179 2271 84 87 7 34 3473 5588 102 164 831 2356 1902 76 61 9 10 6927 2954 693 295 10 70 6248 2316 89 33 11 1999300 13564 46 68 12 153 7963 5858 52 38 13 330 11726 8472 35 26 14 38226446 40829 69 107 15 32 15200 7808 475 244 16 1599 92742 52767 58 33 17881 66957 31410 76 35 18 232 20197 10725 87 46 19 47 2995 1857 64 39 2037 3042 2262 82 61 21 384 17678 10996 46 28 ^(a)K_(M) values of ratnNOS, 1.3 μM; murine iNOS, 8.2 μM; bovine eNOS, 1.7 μM). K_(i) =IC₅₀/(1 + [S]/K_(M)). ^(b)The ratio of K_(i) (eNOS or iNOS) to nNOS

The present invention can also, as would be understood by those skilledin the art, be extended to or include methods using or in conjunctionwith a pharmaceutical composition comprising an inhibitor compound ofthe sort described herein and a physiologically or otherwise suitableformulation. In a some embodiments, the present invention includes oneor more NOS inhibitors, as set forth above, formulated into compositionstogether with one or more physiologically tolerable or acceptablediluents, carriers, adjuvants or vehicles that are collectively referredto herein as carriers. Compositions suitable for such contact oradministration can comprise physiologically acceptable sterile aqueousor nonaqueous solutions, dispersions, suspensions or emulsions. Theresulting compositions can be, in conjunction with the various methodsdescribed herein, for administration or contact with a nitric oxidesynthase. Whether or not in conjunction with a pharmaceuticalcomposition, “contacting” means that a nitric oxide synthase and one ormore inhibitor compounds are brought together for purpose of bindingand/or complexing such an inhibitor compound to the enzyme. Amounts of acompound effective to inhibit a nitric oxide synthase may be determinedempirically, and making such determinations is within the skill in theart. Inhibition or otherwise affecting nitric oxide synthase activityincludes modulation, reduction and/or mitigation, as well as eliminationof NOS activity and/or nitric oxide production.

It is understood by those skilled in the art that dosage amount willvary with the activity of a particular inhibitor compound, diseasestate, route of administration, duration of treatment, and like factorswell-known in the medical and pharmaceutical arts. In general, asuitable dose will be an amount which is the lowest dose effective toproduce a therapeutic or prophylactic effect. If desired, an effectivedose of such a compound, pharmaceutically-acceptable salt thereof, orrelated composition may be administered in two or more sub-doses,administered separately over an appropriate period of time.

Methods of preparing pharmaceutical formulations or compositions includethe step of bringing an inhibitor compound into association with acarrier and, optionally, one or more additional adjuvants oringredients. For example, standard pharmaceutical formulation techniquescan be employed, such as those described in Remington's PharmaceuticalSciences, Mack Publishing Company, Easton, Pa.

Regardless of composition or formulation, those skilled in the art willrecognize various avenues for medicament administration, together withcorresponding factors and parameters to be considered in rendering sucha medicament suitable for administration. Accordingly, with respect toone or more non-limiting embodiments, the present invention provides foruse of one or more neuronal nitric oxide synthase inhibitor compoundsfor the manufacture of a medicament for therapeutic use in the treatmentor prevention of a disease state indicated by nitric oxide production.

EXAMPLES OF THE INVENTION

The following non-limiting examples and data illustrate various aspectsand features relating to the compounds and/or methods of the presentinvention, including the preparation of selective NOS inhibitorcompounds, as available through the synthetic methodologies describedherein. In comparison with the prior art, the present compounds,compositions and methods provide results and data which are surprising,unexpected and contrary thereto. While the utility of this invention isillustrated through the use of several compounds and structural moietieswhich can be incorporated therein, it will be understood by thoseskilled in the art that comparable results are obtainable with variousother structurally analogous compounds and incorporated moieties, as arecommensurate with the scope of this invention.

With reference to the preceding synthetic schemes, the followingexamples provide spectroscopic and related data characterizing theindicated compounds.

Example 1

Compound 1

M.p. 96-98° C. ¹H NMR (500 MHz, CDCl₃) δ 1.88-1.94 (m, 2H), 2.23 (s,3H), 2.90 (t, J=6.5 Hz, 2H), 4.03 (t, J=6.5 Hz, 2H), 4.36 (br, 2H), 4.95(s, 2H), 6.25 (s, 1H), 6.51 (d, J=7.0 Hz, 1H), 6.56-6.58 (m, 2H), 6.69(s, 1H), 7.16 (t, J=8.5 Hz, 1H); ¹³C NMR (125 MHz, CDCl₃) δ 21.1, 33.0,39.3, 65.8, 70.6, 101.7, 106.9, 107.2, 108.0, 112.9, 129.8, 149.7,155.1, 158.1, 159.8, 160.1 ppm; MS (ESI): 288.3 (M+H)⁺. HRMS (ESI):calcd. 288.1712. Found: 288.1717.

Example 2

Compound 2

M.p. 97-99° C. ¹H NMR (500 MHz, CDCl₃) δ 2.23 (s, 6H), 4.41 (br, 2H),4.95 (s, 2H), 5.07 (s, 2H), 6.26 (s, 1H), 6.57 (d, J=7.0 Hz, 1H),6.61-6.63 (m, 2H), 6.67 (s, 1H), 7.18 (t, J=8.0 Hz, 1H), 7.33 (d, J=5.5Hz, 2H), 8.60 (d, J=5.5 Hz, 2H); ¹³C NMR (125 MHz, CDCl₃) δ 21.1, 68.1,70.6, 102.1, 107.4, 107.7, 108.1, 112.9, 130.1, 146.2, 149.7, 150.0,154.8, 158.2, 159.3, 159.9 ppm; MS (ESI): 322.3 (M+H)⁺. HRMS (ESI):calcd. 322.1556. Found: 322.1552 (M+H)⁺.

Example 3

Compound 3

white solid 62 mg. 94%. M.p. 103-105° C. ¹H NMR (500 MHz, DMSO) δ1.67-1.76 (m, 4H), 2.13 (s, 3H), 2.42-2.45 (m, 1H), 2.93 (d, J=11.5 Hz,2H), 3.74 (d, J=6.5 Hz, 2H), 4.83 (s, 2H), 5.82 (s, 2H), 6.18 (s, 1H),6.44 (s, 1H), 6.51 (t, J=7.5 Hz, 1H), 7.13 (t, J=7.5 Hz, 1H), 8.29 (s,2H); ¹³C NMR (125 MHz, CDCl₃) δ 20.7, 29.8, 36.0, 45.7, 70.2, 72.5,101.2, 106.7, 106.8, 107.1, 110.6, 129.8, 147.8, 154.4, 154.5, 159.5,159.9 ppm; MS (ESI): 328.4 (M+H)⁺. HRMS (ESI): calcd. 328.2025. Found:328.2027 (M+H)⁺.

Example 4

Compound 4

M.p. 137-139° C. ¹HNMR (500 MHz, CDCl₃) δ 2.23 (s, 3H), 2.81 (t, J=7.0Hz, 2H), 2.88 (t, J=7.0 Hz, 2H), 3.77 (s, 2H), 4.37 (br, 2H), 4.96 (s,2H), 6.25 (s, 1H), 6.70 (s, 1H), 6.85-6.91 (m, 4H), 6.94 (s, 1H), 6.97(d, J=7.5 Hz, 1H), 7.20-7.24 (m, 2H); ¹³C NMR (125 MHz, CDCl₃) δ 21.14,36.12, 50.08, 53.72, 70.44, 107.98, 112.91+112.94, 113.11, 113.19,114.55, 115.42+115.59, 120.60, 124.36, 124.38, 129.41+129.79, 129.85,141.79, 142.57+142.63, 149.72, 155.14, 158.10, 158.76 ppm. MS (ESI):366.4 (M+H)⁺. HRMS (ESI): calcd. 366.1982. Found: 366.1980 (M+H)⁺.

Example 5

Compound 5

M.p. 128-130° C. ¹HNMR (500 MHz, CDCl₃) δ 2.23 (s, 6H), 2.45 (s, 3H),2.80 (t, J=6.5 Hz, 2H), 2.86 (t, J=6.5 Hz, 2H), 3.79 (s, 2H), 5.00 (s,2H), 6.26 (s, 1H), 6.65 (s, 1H), 6.88-6.91 (m, 2H), 6.96 (d, J=7.0 Hz,1H), 7.22-7.26 (m, 2H), 8.14 (s, 1H), 8.28 (s, 1H); ¹³C NMR (125 MHz,CDCl₃) δ 21.86, 36.03, 50.03, 50.83, 70.57, 108.39, 112.89,113.09+113.25, 115.46+115.63, 121.15, 124.38, 129.90+129.96, 136.22,137.14, 142.01, 142.34+142.40, 149.97, 154.00, 154.87, 155.62, 158.34ppm. MS (ESI): 367.5 (M+H)⁺. HRMS (ESI): calcd. 367.1934. Found:367.1939 (M+H)⁺.

Example 6

Compound 6

M.p. 124-126° C. ¹HNMR (500 MHz, CDCl₃) δ 6.61 (s, 1H), 6.57 (s, 1H),6.23 (s, 2H), 4.43 (q, J=10.0 Hz, J=5.0 Hz, 4H), 4.36 (d, J=5.0 Hz, 4H),4.18-4.16 (m, 1H), 3.69-3.64 (m, 1H), 3.55-3.49 (m, 2H), 3.13-3.11 (m,2H), 2.24 (s, 3H), 2.23 (s, 3H), 2.08 (dd, J=14.0 Hz, J=5.0 Hz, 1H),1.73-1.67 (m, 1H); ¹³C NMR (126 MHz, CDCl₃) δ 158.27, 158.12, 156.39,156.24, 149.58, 149.55, 113.10, 112.97, 107.86, 107.81, 80.30, 73.76,71.63, 56.68, 52.02, 45.99, 34.96, 21.12 ppm; MS (ESI): 358.4 (M+H)⁺.HRMS (ESI): calcd. 358.2243. Found: 358.2234 (M+H)⁺.

Example 7

Compound 7

M.p. 126-128° C. ¹H NMR (500 MHz, CDCl₃) δ 6.60 (s, 1H), 6.59 (s, 1H),6.22 (s, 2H), 4.45 (m, 2H), 4.37 (s, 2H), 4.39-4.35 (m, 1H), 3.78-3.74(m, 1H), 3.65-3.60 (m, 2H), 3.49-3.44 (m, 1H), 3.18 (d, J=10.0 Hz, 1H),2.98-2.94 (m, 1H), 2.21 (s, 6H), 2.18 (dd, J=10.0 Hz, J=5.0 Hz, 1H),1.69-1.65 (m, 1H); ¹³C NMR (126 MHz, CDCl₃) δ 158.17, 158.09, 156.34,156.23, 149.54, 112.97, 112.94, 107.80, 107.75, 80.08, 73.71+73.57,71.61, 57.63, 52.23, 42.91, 35.05, 21.10, 21.08 ppm; MS (ESI): 358.4(M+H)⁺. HRMS (ESI): calcd. 358.2243. Found: 358.2242 (M+H)⁺.

Example 8

Compound 8

M.p. 124-126° C. ¹H NMR (500 MHz, CDCl₃) δ 6.60 (s, 1H), 6.59 (s, 1H),6.22 (s, 2H), 4.45 (m, 2H), 4.37 (s, 2H), 4.39-4.35 (m, 1H), 3.78-3.74(m, 1H), 3.65-3.60 (m, 2H), 3.49-3.44 (m, 1H), 3.18 (d, J=10.0 Hz, 1H),2.98-2.94 (m, 1H), 2.21 (s, 6H), 2.18 (dd, J=10.0 Hz, J=5.0 Hz, 1H),1.69-1.65 (m, 1H); ¹³C NMR (126 MHz, CDCl₃) δ 158.17, 158.09, 156.34,156.23, 149.54, 112.97, 112.94, 107.80, 107.75, 80.08, 73.71+73.57,71.61, 57.63, 52.23, 42.91, 35.05, 21.10, 21.08 ppm; MS (ESI): 358.4(M+H)⁺. HRMS (ESI): calcd. 358.2243. Found: 358.2236 (M+H)⁺.

Example 9

Compound 9

M.p. 126-128° C. ¹H NMR (500 MHz, CDCl₃) δ 6.61 (s, 1H), 6.57 (s, 1H),6.23 (s, 2H), 4.43 (q, J=10.0 Hz, J=5.0 Hz, 4H), 4.36 (d, J=5.0 Hz, 4H),4.18-4.16 (m, 1H), 3.69-3.64 (m, 1H), 3.55-3.49 (m, 2H), 3.13-3.11 (m,2H), 2.24 (s, 3H), 2.23 (s, 3H), 2.08 (dd, J=14.0 Hz, J=5.0 Hz, 1H),1.73-1.67 (m, 1H); ¹³C NMR (126 MHz, CDCl₃) δ 158.29, 158.18, 156.37,156.28, 149.54, 149.51, 112.99, 112.82, 107.88, 107.82, 80.20, 73.67,71.58, 56.66, 51.88, 45.90, 34.89, 21.10 ppm; MS (ESI): 358.4 (M+H)⁺.HRMS (ESI): calcd. 358.2234. Found: 358.2238.

Example 10

Compound 10

M.p. 136-138° C. ¹H NMR (500 MHz, CDCl₃) δ 6.49 (s, 2H), 6.26 (s, 2H),4.46-4.30 (m, 6H), 3.78-3.67 (m, 6H), 3.43-3.28 (m, 2H), 2.94-2.78 (m,1H), 2.22 (s, 3H), 2.20 (s, 3H), 2.18-2.12 (m, 1H); ¹³C NMR (126 MHz,CDCl₃) δ 158.46, 158.29, 154.15, 153.68, 150.57, 150.38, 128.43,113.28+113.14, 113.06+112.95, 108.88, 108.74, 78.30, 72.88, 70.68,70.55, 69.81, 57.37, 50.06, 45.30, 36.76, 28.48, 21.17, 21.15 ppm; MS(ESI): 402.7 (M+H)⁺. calcd. 402.2505. Found: 402.2492 (M+H)⁺.

Example 11

Compound 11

pale yellow solid 15 mg, 73%. M.p. 139-141° C. ¹H NMR (500 MHz, CDCl₃) δ6.55-6.51 (m, 2H), 6.21 (s, 2H), 4.45-4.35 (m, 4H), 4.22 (s, 1H),3.72-3.59 (m, 8H), 3.37-3.08 (m, 2H), 2.22 (s, 3H), 2.21 (s, 3H),2.05-2.02 (m, 1H); ¹³C NMR (126 MHz, CDCl₃) δ 158.55, 158.37, 155.38,153.20, 149.93, 149.77, 113.16, 112.92, 108.25, 108.23, 79.34+78.92,73.54, 72.43, 71.09, 70.53, 69.90, 57.04+56.82, 53.47, 50.99, 50.47,34.20, 29.43, 21.12 ppm; MS (ESI): 402.8 (M+H)⁺. calcd. 402.2505. Found:402.2498 (M+H)⁺.

Example 12

Compound 12

M.p. 137-139° C. ¹H NMR (500 MHz, CDCl₃) δ 7.29-7.26 (m, 1H), 7.10-7.06(m, 2H), 6.96 (t, J=7.0 Hz, 1H), 6.55 (s, 1H), 6.22 (s, 1H), 4.55 (s,2H), 4.41-4.39 (m, 2H), 3.68-3.56 (m, 6H), 3.50-3.47 (m, 2H), 3.47-3.10(m, 2H), 2.22 (s, 3H), 2.06 (dd, J=6.5 Hz, J=13.5 Hz, 1H), 1.74-1.71 (m,1H); ¹³C NMR (126 MHz, CDCl₃) δ 163.91, 161.95, 158.30, 149.86,140.98+140.92, 129.91+129.84, 122.98, 114.50+114.42+114.33+114.24,112.91, 108.12, 79.88, 73.20+73.11, 72.36, 71.06, 70.54, 69.62+69.56,56.57, 51.42, 34.70+34.60, 21.10 ppm; MS (ESI): 390.7 (M+H)⁺. HRMS (ESI)calcd. 390.2193. Found: 390.2198 (M+H)⁺.

Example 13

Compound 13

M.p. 139-141° C. ¹H NMR (500 MHz, CDCl₃) δ 7.29-7.26 (m, 1H), 7.08 (d,J=7.5 Hz, 1H), 7.05 (d, J=7.5 Hz, 1H), 6.96 (t, J=7.5 Hz, 1H), 6.52 (s,1H), 6.21 (s, 1H), 4.54 (s, 4H), 4.41-4.20 (m, 2H), 3.70 (br, 1H),3.69-3.63 (m, 4H), 3.58-3.53 (m, 1H), 3.26 (d, J=11.0 Hz, 1H), 3.02-2.99(m, 1H), 2.21 (s, 3H), 2.17-2.15 (m, 2H); ¹³C NMR (126 MHz, CDCl₃) δ163.88, 161.93, 158.32, 155.64+155.50, 149.85, 140.94+140.88,129.91+129.84, 123.02, 114.49+114.45+114.32+114.28, 112.90+112.84,108.13, 79.38, 73.07, 72.44, 71.05, 70.50, 69.62+69.55, 61.91, 57.02,51.08, 34.59, 21.08 ppm; MS (ESI): 390.7 (M+H)⁺. calcd. 390.2193. Found:390.2192 (M+H)⁺.

Example 14

Compound 14

M.p. 133-135° C. ¹H NMR (500 MHz, CDCl₃) δ 6.50 (s, 1H), 6.45 (s, 1H),6.20 (s, 2H), 4.71 (br, 2H), 4.63-4.44 (m, 2H), 4.41-4.37 (m, 2H),3.76-3.65 (m, 3H), 3.19-2.99 (m, 2H), 2.78 (br, 4H), 2.19 (s, 6H); ¹³CNMR (126 MHz, CDCl₃) δ 158.77, 155.62, 155.41, 149.57, 149.43, 112.96,112.63, 108.30, 108.06, 77.95, 73.65, 72.28, 71.59, 41.39, 21.02 ppm; MS(ESI): 332.7 (M+H)⁺. HRMS (ESI): calcd. 332.2087. Found: 332.2081(M+H)⁺.

Example 15

Compound 15

M.p. 128-130° C. ¹H NMR (500 MHz, CDCl₃) δ 6.61 (s, 1H), 6.55 (s, 1H),6.22 (s, 2H), 6.80 (s, 1H), 4.63-4.50 (m, 4H), 4.46 (d, J=6.5 Hz, 1H),3.67 (s, 2H), 2.93-2.89 (m, 2H), 2.21 (s, 6H); ¹³C NMR (126 MHz, CDCl₃)δ 158.34, 158.32, 156.30, 156.00, 149.52, 149.47, 113.15, 112.91,107.95, 107.87, 79.50, 73.93, 72.58, 71.07, 42.78, 21.07 ppm; MS (ESI):332.3 (M+H)⁺. HRMS: (ESI): calcd. 332.2087. Found: 332.2082 (M+H)⁺.

Example 16

Compound 16

M.p. 133-135° C. ¹H NMR (500 MHz, CDCl₃) δ 6.57 (s, 1H), 6.50 (s, 1H),6.22 (s, 1H), 6.19 (s, 1H), 4.66 (d, J=12.5 Hz, 1H), 4.49 (d, J=12.5 Hz,1H), 4.45 (s, 3H), 3.79-3.76 (m, 1H), 3.01-2.92 (m, 3H), 2.22 (s, 3H),2.19 (s, 3H), 1.85-1.81 (m, 2H); ¹³C NMR (126 MHz, CDCl₃) δ 158.55,158.28, 156.15, 156.10, 149.49, 149.47, 112.93, 112.87, 108.00, 107.85,77.89, 74.11, 72.83, 72.31, 38.50, 30.99, 21.11, 21.06 ppm. MS (ESI):346.7 (M+H)⁺. HRMS: (ESI): calcd. 346.2243. Found: 332.2250 (M+H)⁺.

Example 17

Compound 17

M.p. 137-140° C. ¹H NMR (500 MHz, CDCl₃) δ 6.57 (s, 1H), 6.50 (s, 1H),6.22 (s, 1H), 6.19 (s, 1H), 4.66 (d, J=12.5 Hz, 1H), 4.49 (d, J=12.5 Hz,1H), 4.45 (s, 3H), 3.79-3.76 (m, 1H), 3.01-2.92 (m, 3H), 2.22 (s, 3H),2.19 (s, 3H), 1.85-1.81 (m, 2H); ¹³C NMR (126 MHz, CDCl₃) δ 158.65,158.48, 156.17, 156.13, 149.49, 149.47, 112.93, 112.87, 108.00, 107.89,77.88, 74.11, 72.83, 72.31, 38.50, 30.99, 21.11, 21.06 ppm. MS (ESI):346.5 (M+H)⁺. HRMS: (ESI): calcd. 346.2243. Found: 346.2245 (M+H)⁺.

Example 18

Compound 18

M.p. 134-136° C. ¹H NMR (500 MHz, CDCl₃) δ 6.50 (s, 1H), 6.46 (s, 1H),6.21 (s, 1H), 6.22 (s, 1H), 4.52 (d, J=12.5 Hz, 1H), 4.42-4.37 (m, 3H),3.67-3.64 (m, 1H), 3.62-3.57 (m, 2H), 3.08 (dd, J=10.0 Hz, J=5.0 Hz,1H), 2.88 (dd, J=10.0 Hz, J=5.0 Hz, 1H), 2.20 (s, 3H), 2.18 (s, 3H),1.96-1.1.88 (m, 2H); ¹³C NMR (126 MHz, CDCl₃) δ 158.64, 158.55, 155.89,155.77, 149.62, 149.46, 112.99, 112.88, 108.24, 108.05, 73.72, 72.31,66.73, 43.62, 32.88, 21.06 ppm. MS (ESI): 346.7 (M+H)⁺. HRMS: (ESI):calcd. 346.2243. Found: 346.2241 (M+H)⁺.

Example 19

Compound 19

M.p. 127-129° C. ¹H NMR (500 MHz, CDCl₃) δ 6.48 (s, 1H), 6.46 (s, 1H),6.22 (s, 1H), 6.21 (s, 1H), 4.59 (d, J=10.0 Hz, 1H), 4.48-4.35 (m, 3H),3.69-3.62 (m, 2H), 3.07-3.05 (m, 1H), 2.76-2.72 (m, 1H), 2.21 (s, 6H),1.23 (d, J=5.0 Hz, 3H); ¹³C NMR (126 MHz, CDCl₃) δ 158.52, 158.37,156.21, 156.76, 149.62, 149.59, 112.73, 112.52, 108.09, 107.92, 76.32,73.36, 71.17, 56.20, 50.81, 21.08, 21.05, 16.26 ppm. MS (ESI): 346.2(M+H)⁺. HRMS: (ESI): calcd. 346.2243. Found: 332.2241, (M+H)⁺.

Example 20

Compound 20

M.p. 125-127° C. ¹H NMR (500 MHz, CDCl₃) δ 6.48 (s, 1H), 6.46 (s, 1H),6.22 (s, 1H), 6.21 (s, 1H), 4.59 (d, J=10.0 Hz, 1H), 4.48-4.35 (m, 3H),3.69-3.62 (m, 2H), 3.07-3.05 (m, 1H), 2.76-2.72 (m, 1H), 2.21 (s, 6H),1.23 (d, J=5.0 Hz, 3H); ¹³C NMR (126 MHz, CDCl₃) δ 158.52, 158.37,156.21, 156.76, 149.62, 149.59, 112.73, 112.52, 108.09, 107.92, 76.32,73.36, 71.17, 56.20, 50.81, 21.08, 21.05, 16.26 ppm. MS (ESI): 346.2(M+H)⁺. HRMS: (ESI): calcd. 346.2243. Found: 332.2239 (M+H)⁺.

Example 21

Compound 21

M.p. 129-132° C. ¹H NMR (500 MHz, CDCl₃) δ 6.54 (s, 2H), 6.22 (s, 1H),6.21 (s, 1H), 4.53-4.35 (m, 5H), 3.73-3.70 (m, 2H), 3.51-3.47 (m, 1H),2.21 (s, 6H), 1.23 (d, J=5.0 Hz, 3H); ¹³C NMR (126 MHz, CDCl₃) δ 158.28,158.19, 156.60, 156.51, 149.53, 149.51, 112.90, 112.76, 107.88, 107.79,73.77, 72.29, 71.35, 54.65, 21.10, 21.08, 15.29 ppm. MS (ESI): 346.2(M+H)⁺. HRMS: (ESI): calcd. 346.2243 (M+H)⁺. Found: 332.2243, (M+H)⁺.

We claim:
 1. A compound selected from compounds of a formula

and salts thereof.
 2. The compound of claim 1 selected from cis andtrans diastereomers.
 3. The compound of claim 1 wherein said compound isan ammonium salt.
 4. The compound of claim 3 wherein said ammonium salthas a counter ion that is a conjugate base of a protic acid.